Kinetics of drug selection systems in mouse embryonic stem cells

Yuhki Nakatake, Setsuko Fujii, Shinji Masui, Toshimi Sugimoto, Satomi Torikai-Nishikawa, Kenjiro Adachi, Hitoshi Niwa

研究成果: Article査読

16 被引用数 (Scopus)

抄録

Background: Stable expression of transgenes is an important technique to analyze gene function. Various drug resistance genes, such as neo, pac, hph, zeo, bsd, and hisD, have been equally used as selection markers to isolate a transfectant without considering their dose-dependent characters.Results: We quantitatively measured the variation of transgene expression levels in mouse embryonic stem (mES) cells, using a series of bi-cistronic expression vectors that contain Egfp expression cassette linked to each drug resistant gene via IRES with titration of the selective drugs, and found that the transgene expression levels achieved in each system with this vector design are in order, in which pac and zeo show sharp selection of transfectants with homogenously high expression levels. We also showed the importance of the choice of the drug selection system in gene-trap or gene targeting according to this order.Conclusions: The results of the present study clearly demonstrated that an appropriate choice of the drug resistance gene(s) is critical for a proper design of the experimental strategy.

本文言語English
論文番号64
ジャーナルBMC Biotechnology
13
DOI
出版ステータスPublished - 2013 8月 7
外部発表はい

ASJC Scopus subject areas

  • バイオテクノロジー

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